vasoactive-intestinal-peptide and Peritonitis

To facilitate the targeting to inflammation sites of small anti-inflammatory peptides, with short half-lives, by fusion with the latency-associated peptide (LAP) of transforming growth factor β1 through a cleavable matrix metalloproteinase (MMP) linker. This design improves efficacy, overcoming the limitations to their clinical use.. We generated latent forms of vasoactive intestinal peptide (VIP), α-melanocyte-stimulating hormone (MSH) and γ(3)MSH by fusion to LAP through an MMP cleavage site using recombinant DNA technology. The biological activities of these latent therapeutics were studied in vivo using monosodium urate (MSU)-induced peritonitis and collagen-induced arthritis (CIA) models. We assessed gene therapy and purified protein therapy.. The recruitment of the polymorphonuclear cells induced by MSU injection into mouse peritoneal cavity was reduced by 35% with γ(3)MSH (1 nmol), whereas administration of a much lower dose of purified latent LAP-MMP-γ(3)MSH (0.03 nmol) attenuated leucocyte influx by 50%. Intramuscular gene delivery of plasmids coding LAP-MMP-VIP and LAP-MMP-αMSH at disease onset reduced the development of CIA compared with LAP-MMP, which does not contain any therapeutic moiety. Histological analysis confirmed a significantly lower degree of inflammation, bone and cartilage erosion in groups treated with LAP-MMP-VIP or LAP-MMP-αMSH. Antibody titres to collagen type II and inflammatory cytokine production were also reduced in these two groups.. Incorporation of small anti-inflammatory peptides within the LAP shell and delivered as recombinant protein or through gene therapy can control inflammatory and arthritic disease. This platform delivery can be developed to control human arthritides and other autoimmune diseases.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Arthritis, Experimental; Cytokines; Drug Delivery Systems; Drug Design; Drug Evaluation, Preclinical; Genetic Therapy; Half-Life; Male; Melanocyte-Stimulating Hormones; Mice; Mice, Inbred DBA; Peptide Fragments; Peritonitis; Recombinant Fusion Proteins; Tissue Distribution; Transforming Growth Factor beta; Treatment Outcome; Vasoactive Intestinal Peptide

Vasoactive intestinal peptide (VIP) is a neuropeptide released from the autonomic nerves exerting multiple antiinflammatory effects. The aim of the present study was to investigate the impact of severe sepsis and hemofiltration in two settings on plasma and tissue concentrations of VIP in a porcine model of sepsis. Thirty-two pigs were divided into 5 groups: 1) control group; 2) control group with conventional hemofiltration; 3) septic group; 4) septic group with conventional hemofiltration; 5) septic group with high-volume hemofiltration. Sepsis induced by faecal peritonitis continued for 22 hours. Hemofiltration was applied for the last 10 hours. Hemodynamic, inflammatory and oxidative stress parameters (heart rate, mean arterial pressure, cardiac output, systemic vascular resistance, plasma concentrations of tumor necrosis factor-alpha, interleukin-6, thiobarbituric acid reactive species, nitrate + nitrite, asymmetric dimethylarginine) and the systemic VIP concentrations were measured before faeces inoculation and at 12 and 22 hours of peritonitis. VIP tissue levels were determined in the left ventricle, mesenteric and coronary arteries. Sepsis induced significant increases in VIP concentrations in the plasma and mesenteric artery, but it decreased peptide levels in the coronary artery. Hemofiltration in both settings reduced concentrations of VIP in the mesenteric artery. In severe sepsis, VIP seems to be rapidly depleted from the coronary artery and, on the other hand, upregulated in the mesenteric artery. Hemofiltration in both settings has a tendency to drain away these upregulated tissue stores which could result in the limited secretory capacity of the peptide.

Topics: Animals; Coronary Vessels; Female; Hemofiltration; Male; Mesenteric Arteries; Oxidative Stress; Peritonitis; Sepsis; Swine; Vasoactive Intestinal Peptide

Gastrointestinal motility is reduced during sepsis but the pathomechanism involved is poorly understood. We investigated the expression of substance P (SP) and vasoactive intestinal peptide (VIP) in the myenteric plexus during peritonitis in human small bowel.. Tissue samples of the small bowel were gathered from healthy patients and from patients with peritonitis. Immunohistochemistry for myeloperoxidase (MPO), SP, and VIP was performed in whole mount sections. To determine the level of inflammation, MPO-positive cells were counted in the circular muscle layer. SP and VIP immunoreactivity was analyzed in myenteric plexus neurons. The area of positive immunoreactivity for either neuropeptide within the plexus was analyzed and set in relation to the total area of the plexus and consecutively expressed as percentage.. During peritonitis, MPO-positive cells significantly increased by approximately fourfold as compared to healthy tissue. The immunoreactivity for SP was significantly reduced by approximately 80% in myenteric plexus neurons during peritonitis. In contrast, the immunoreactivity for VIP significantly increased by nearly twofold during peritonitis.. During peritonitis, the inflammatory reaction within the gut is increased. The neuropeptide expression in myenteric plexus neurons was observed as shifting towards increased expression of VIP, known to inhibit intestinal motility, and towards decreased expression of the prokinetic neuropeptide SP.

Topics: Adult; Aged; Aged, 80 and over; Female; Gastrointestinal Motility; Humans; Intestine, Small; Male; Middle Aged; Myenteric Plexus; Neurons; Peritonitis; Peroxidase; Substance P; Vasoactive Intestinal Peptide

Inflammatory chemokines recruit various populations of immune cells that initiate and maintain the inflammatory response against foreign Ags. Although such a response is necessary for the elimination of the Ag, the inflammation has to be eventually resolved in a healthy organism. Neuropeptides such as vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP), released after antigenic stimulation, contribute to the termination of an inflammatory response primarily by inhibiting the production of proinflammatory cytokines. Here we investigated the effects of VIP and PACAP on chemokine production. We report that VIP and PACAP inhibit the expression of the macrophage-derived CXC chemokines macrophage inflammatory protein-2 and KC (IL-8), and of the CC chemokines MIP-1alpha, MIP-1beta, monocyte chemoattractant protein 1, and RANTES in vivo and in vitro. The inhibition of chemokine gene expression correlates with an inhibitory effect of VIP/PACAP on NF-kappaB binding and transactivating activity. The VIP/PACAP inhibition of both chemokine production and of NF-kappaB binding and transactivating activity is mediated through the specific VIP receptor VPAC1, and involves both cAMP-dependent and -independent intracellular pathways. In an in vivo model of acute peritonitis, the inhibition of chemokine production by VIP/PACAP leads to a significant reduction in the recruitment of polymorphonuclear cells, macrophages, and lymphocytes into the peritoneal cavity. These findings support the proposed role of VIP and PACAP as key endogenous anti-inflammatory agents and describe a novel mechanism, i.e., the inhibition of the production of macrophage-derived chemokines.

Topics: Acute Disease; Animals; Cell Line; Cell Migration Inhibition; Cells, Cultured; Chemokines; Chemokines, CC; Chemokines, CXC; Disease Models, Animal; Gene Expression Regulation; Injections, Intraperitoneal; Interleukin-10; Intracellular Fluid; Lipopolysaccharides; Macrophages, Peritoneal; Male; Mice; Mice, Inbred BALB C; Neuropeptides; NF-kappa B; Peritonitis; Pituitary Adenylate Cyclase-Activating Polypeptide; Protein Binding; Receptors, Vasoactive Intestinal Peptide; Receptors, Vasoactive Intestinal Polypeptide, Type I; RNA, Messenger; Signal Transduction; Vasoactive Intestinal Peptide

Inflammatory chemokines recruit immune cells which initiate and maintain the inflammatory response. Although such a response is necessary for the elimination of the antigen, the inflammation has to be eventually resolved. Peptides such as vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP), released following antigenic stimulation, contribute to the termination of an inflammatory response primarily by inhibiting the production of proinflammatory cytokines. Here we investigated the effects of VIP and PACAP on chemokine production. We report that VIP and PACAP inhibit the expression of the macrophage-derived CXC chemokines MIP-2 and KC (IL-8), and of the CC chemokines MIP-1a, MIP-1b, MCP-1 and RANTES in vivo and in vitro. The decrease of chemokine gene expression correlates with an inhibitory effect of VIP/PACAP on NFkB binding. In an in vivo model of acute peritonitis, the inhibition of chemokine production by VIP/PACAP leads to a significant reduction in the recruitment of PMNs, macrophages and lymphocytes into the peritoneal cavity. These findings support the proposed role of VIP and PACAP as key endogenous anti-inflammatory agents, and describe a novel mechanism, i.e., the inhibition of the production of macrophage-derived chemokines.

Topics: Animals; Cell Fractionation; Cells, Cultured; Chemokines; Disease Models, Animal; Leukocytes; Lipopolysaccharides; Macrophage Activation; Macrophages, Peritoneal; Mice; Mice, Inbred BALB C; Neuropeptides; NF-kappa B; Peritonitis; Pituitary Adenylate Cyclase-Activating Polypeptide; Vasoactive Intestinal Peptide

Peritonitis induced by serosal application of 0.1 M hydrochloric acid causes net fluid secretion via the enteric nervous system. The aim of the present study was to investigate the roles of vasoactive intestinal peptide (VIP) and tachykinins in this reflex(es). The release of tachykinins (substance P [SP], neurokinin A [NKA], neuropeptide K [NPK]) and VIP into the mesenteric circulation, net fluid transport, intestinal blood flow and sometimes motility were recorded simultaneously in extrinsically denervated jejunal segments of the cat in vivo. The release of both VIP and NKA was increased upon application of HCl to the cat jejunal serosa. Tetrodotoxin, hexamethonium and methionine enkephalin inhibited both the induced VIP release and the secretory response. The increased release of NKA was unaffected by hexamethonium. We propose that the intramural secretory reflex evoked by acid application of the serosa consists of an 'afferent' tachykinin neuron, a cholinergic interneuron and an 'efferent' VIPergic neuron innervating the secretory enterocytes.

Topics: Animals; Biological Transport; Body Fluids; Cats; Enkephalin, Methionine; Female; Ganglionic Blockers; Hexamethonium; Hexamethonium Compounds; Intestine, Small; Male; Peritonitis; Reflex; Tachykinins; Tetrodotoxin; Vasoactive Intestinal Peptide

Rats were subjected to laparotomy, to pancreatectomy, to gastroenterectomy (control groups), or to these procedures plus a septic challenge by instillation of 0.1 ml feces intraperitoneally (experimental groups). In the laparotomized controls plasma somatostatin values were significantly higher in samples from the portal vein than from the upper inferior caval vein. After both pancreatectomy and gastroenterectomy a significant fall in plasma somatostatin values was observed, and there was no significant difference between samples taken from the portal vein and the systemic circulation. An intraperitoneal septic challenge elicited a significant rise in portal plasma somatostatin in laparotomized rats, whereas this increase did not occur in pancreatectomized and gastroenterectomized animals, supporting the notion that plasma somatostatin originates from the pancreas and/or the gastrointestinal tract during septic peritonitis. No differences were detected in plasma vasoactive intestinal polypeptide (VIP) values in animals from the three control groups. However, both in laparotomized and in pancreatectomized septic animals a significant rise in plasma VIP was demonstrated in samples from the portal vein. By contrast, no such increase was observed in gastroenterectomized septic rats. Thus, the gastrointestinal tract seems to be the major source of circulating VIP during fecal peritonitis in the rat.

Topics: Animals; Feces; Gastrectomy; Intestines; Male; Pancreatectomy; Peritonitis; Rats; Rats, Inbred Strains; Somatostatin; Vasoactive Intestinal Peptide

Vasoactive intestinal polypeptide (VIP) and somatostatin were measured during endogenous gram-negative peritonitis and septicaemia in rats. Both peptides were found to increase in blood, but not in peritoneal fluid. The VIP values coincided with the levels of endotoxin and bacterial counts. However, if the development of profound shock was prevented by intravenous fluid supply, scarcely any changes in plasma VIP or somatostatin were found. Somatostatin is known to inhibit VIP. Our findings suggested breakdown of this regulatory inhibition in lethal gram-negative sepsis. They also supported the concept that specific release of the peptides takes place, not merely passive diffusion from injured cells.

Topics: Animals; Endotoxins; Gram-Negative Bacteria; Male; Models, Biological; Peritoneum; Peritonitis; Rats; Rats, Inbred Strains; Sepsis; Somatostatin; Vasoactive Intestinal Peptide